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- Synthesis of pinene in the industrial strain Candida glycerinogenes by modification of its mevalonate pathway
- Tengfei Ma , Hong Zong , Xinyao Lu , Bin Zhuge
- J. Microbiol. 2022;60(12):1191-1200. Published online October 24, 2022
- DOI: https://doi.org/10.1007/s12275-022-2344-0
- 319 View
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Abstract
PDF - Terpenes have many applications and are widely found in nature, but recent progress in synthetic biology has enabled the use of microorganisms as chassis cells for the synthesis of these compounds. Candida glycerinogenes (C. glycerinogenes) is an industrial strain that may be developed as a chassis for the synthesis of terpenes since it has a tolerance to hyperosmolality and high sugar, and has a complete mevalonate (MVA) pathway. However, monoterpenes such as pinene are highly toxic, and the tolerance of C. glycerinogenes to pinene was investigated. We also measured the content of mevalonate and squalene to evaluate the strength of the MVA pathway. To determine terpene synthesis capacity, a pathway for the synthesis of pinene was constructed in C. glycerinogenes. Pinene production was improved by overexpression, gene knockdown and antisense RNA inhibition. Pinene production was mainly enhanced by strengthening the upstream MVA pathway and inhibiting the production of by-products from the downstream pathway. With these strategies, yield could be increased by almost 16 times, to 6.0 mg/L. Overall, we successfully constructed a pinene synthesis pathway in C. glycerinogenes and enhanced pinene production through metabolic modification.
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Citations
Citations to this article as recorded by
- Recent advances in genome mining and synthetic biology for discovery and biosynthesis of natural products
Mingpeng Wang, Lei Chen, Zhaojie Zhang, Qinhong Wang
Critical Reviews in Biotechnology.2025; 45(1): 236. CrossRef - Engineering a complete mevalonate pathway in Chlamydomonas reinhardtii for enhanced isoprenoid production
Jingkai Wang, Muhammad Anwar, Jiancheng Li, Lin Dan, Bin Jia, Zhangli Hu
Algal Research.2025; 88: 103987. CrossRef - Two-Phase Fermentation Systems for Microbial Production of Plant-Derived Terpenes
Tuo Li, Ximeng Liu, Haoyu Xiang, Hehua Zhu, Xuan Lu, Baomin Feng
Molecules.2024; 29(5): 1127. CrossRef - Acetic acid stress and utilization synergistically enhance squalene biosynthesis in Candida glycerinogenes
Zhenzhen You, Xueqing Du, Hong Zong, Xinyao Lu, Bin Zhuge
Biochemical Engineering Journal.2024; 210: 109413. CrossRef - Recent developments in enzymatic and microbial biosynthesis of flavor and fragrance molecules
Roman M. Dickey, Madan R. Gopal, Priyanka Nain, Aditya M. Kunjapur
Journal of Biotechnology.2024; 389: 43. CrossRef - Recent Advances and Multiple Strategies of Monoterpenoid Overproduction in Saccharomyces cerevisiae and Yarrowia lipolytica
Dong-Xun Li, Qi Guo, Yu-Xin Yang, Shun-Jie Jiang, Xiao-Jun Ji, Chao Ye, Yue-Tong Wang, Tian-Qiong Shi
ACS Synthetic Biology.2024; 13(6): 1647. CrossRef - Gene Editing of Candida glycerinogenes by Designed Toxin–Antitoxin Cassette
Wen Lv, Xinyao Lu, Bin Zhuge, Hong Zong
ACS Synthetic Biology.2024; 13(3): 816. CrossRef - Candida glycerinogenes-Promoted α-Pinene and Squalene Co-production Strategy Based on α-Pinene Stress
Tengfei Ma, Hong Zong, Xinyao Lu, Bin Zhuge
Journal of Agricultural and Food Chemistry.2023; 71(13): 5250. CrossRef
- Recent advances in genome mining and synthetic biology for discovery and biosynthesis of natural products
- Screening and identification of Aspergillus activity against Xanthomonas oryzae pv. oryzae and analysis of antimicrobial components
- Bei Jiang , Zhiying Wang , Chuxuan Xu , Weijia Liu , Donghua Jiang
- J. Microbiol. 2019;57(7):597-605. Published online June 27, 2019
- DOI: https://doi.org/10.1007/s12275-019-8330-5
- 267 View
- 0 Download
- 12 Web of Science
- 11 Crossref
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Abstract
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To screen for Aspergillus activity against Xanthomonas oryzae
pv. oryzae and analyse the antimicrobial components
involved, 60 Aspergillus spp. were isolated and purified from
fruits, soil and other habitats. As-75, an Aspergillus strain that
can antagonize Xanthomonas oryzae pv. oryzae, was identified
based on the zone of inhibition formed during co-culture.
According to morphological, ITS rDNA gene sequencing
and phylogenetic tree results, the strain showed close
homology to Aspergillus sclerotiorum. The biochemical characterization
tests showed that the fermentation broth of strain
As-75 exhibited a high capacity for environmental adaptation.
The results of the antimicrobial spectrum experiments demonstrated
that As-75 exhibited fairly strong antagonistic activity
against five plant pathogenic fungi and six plant pathogenic
bacteria in vitro. The fermentation broth of strain As-75
displayed maximum stability under fluorescent illumination
at temperatures below 60°C at pH 6.5. A substance with antagonistic
activity was obtained from strain As-75 via fractional
extraction, silica gel column chromatography and thinlayer
chromatography. Through mass spectrometry, nuclear
magnetic resonance and electrospray ionization mass spectrometry
(ESI-MS) analyses, the target compound was identified
as (2Z)-2-butenedioic acid-2-(1-methylethenyl)-4-methyl
ester; its molecular weight of 170.06 daltons and formula
of C8H10O4 identify it as a novel compound. Trials of
the preventative and curative effects demonstrated that compound
S1 exhibited a better control efficiency than the control
against rice bacterial blight. Additionally, the M1 processing
method
was better, and the efficiency of compound S1 in preventing rice bacterial blight in six rice varieties, TN1, IR24, ZF802, Zhonghua 11, Wuyunjing 21, and Nipponbare, was 78.3%, 77.5%, 74.2%, 75.3%, 70.9%, and 72.1%, respectively. -
Citations
Citations to this article as recorded by- Screening and identification of Aspergillus sclerotiorum with activity against Metschnikowia bicuspidata and initial application on "milky disease" in Eriocheir sinensis
Senting Pu, Zhouling Chen, Dong Sheng, Yunmeng Shan, Peilin Zhou, Xinran Shi, Kexin Hao, Shigen Ye
Aquaculture.2025; 595: 741653. CrossRef - Isolation and anti-Xanthomonas citri activity of unguinol produced by Aspergillus unguis CBMAI 2140
Eduarda Araujo dos Santos, Vítor Rodrigues Marin, Derlene Atilli de Angelis, Henrique Ferreira, Daiane Cristina Sass
Letters in Applied Microbiology.2025;[Epub] CrossRef -
Application and antagonistic mechanisms of atoxigenic
Aspergillus
strains for the management of fungal plant diseases
Suyan Wang, Yanxia Wang, Xinchi Shi, Daniela D. Herrera-Balandrano, Xin Chen, Fengquan Liu, Pedro Laborda, Irina S. Druzhinina
Applied and Environmental Microbiology.2024;[Epub] CrossRef - Screening of indigenous entomopathogenic fungal isolates on plant parasitic nematodes in China
Ming Fang, Jie Sun, Ailing Wang, Hongbo Tang, Lei Wang, Xianqin Wei, Weibin Ruan
European Journal of Plant Pathology.2024; 169(4): 787. CrossRef - Synergy in Rice Immunity: Exploring Strategies of Coordinated Disease Defense Through Receptor-Like Kinases and Receptor- Like Cytoplasmic Kinases
Mengtian Pei, Yingying Cao, Xuze Xie, Ying Cao, Jia Chen, Xi Zhang, Zonghua Wang, Guodong Lu, Shenghang Zhang
Rice Science.2024; 31(6): 643. CrossRef - Antimicrobial cyclic lipopeptides from Bacillus mojovensis B1302 against wheat root rot
Yanjie Yi, Shijie Liu, Shihao Ren, Yunpeng Shen, Xinyue Lin, Jia Shi, Kang Wang, Changfu Zhang
Rhizosphere.2024; 32: 100963. CrossRef - Screening<italic> Streptomyces </italic>against <italic>Xanthomonas axonopodis</italic> pv<italic>. glycines</italic> and study of growth-promoting and biocontrol effect
Xuan-Dong WANG, Sun-Yu-Yue YANG, Run-Jie GAO, Jun-Jie YU, Dan-Pei ZHENG, Feng NI, Dong-Hua JIANG
Acta Agronomica Sinica.2022; 48(6): 1546. CrossRef - Effect of microwave radiation combined with cellulase treatment of soybean residue on the culture of Aspergillus oryzae
Huaixiang Tian, Yao Liu, Li Li, Chen Chen, Haiyan Yu, Xinxin Ma, Juan Huang, Xinman Lou, Haibin Yuan
Food Bioscience.2022; 50: 101988. CrossRef - Compound fermentation supernatants of antagonistic bacteria control Rhizoctonia cerealis and promote wheat growth
Yanjie Yi, Yang Liu, Pengyu Luan, Zhipeng Hou, Yanhui Yang, Ruifang Li, Zhenpu Liang, Xiaoxia Zhang, Shulei Liu
Egyptian Journal of Biological Pest Control.2022;[Epub] CrossRef - Screening of antagonistic bacteria against the blue mold of citrus fruit from soil by a new parallel screening method without prior isolation of single strains
Zhenzhen Sun, Tingting Liu, Zhe Liu, Chaozhen Zeng, Zhixiang Liu
Biological Control.2022; 176: 105066. CrossRef - Berkchaetoazaphilone B has antimicrobial activity and affects energy metabolism
Xudong Ouyang, Jelmer Hoeksma, Gisela van der Velden, Wouter A. G. Beenker, Maria H. van Triest, Boudewijn M. T. Burgering, Jeroen den Hertog
Scientific Reports.2021;[Epub] CrossRef
- Screening and identification of Aspergillus sclerotiorum with activity against Metschnikowia bicuspidata and initial application on "milky disease" in Eriocheir sinensis
- NMR-based metabolomics reveals the metabolite profiles of Vibrio parahaemolyticus under ferric iron stimulation
- Jun Zhou , Chenyang Lu , Dijun Zhang , Chennv Ma , Xiurong Su
- J. Microbiol. 2017;55(8):628-634. Published online July 28, 2017
- DOI: https://doi.org/10.1007/s12275-017-6551-z
- 276 View
- 0 Download
- 6 Crossref
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Abstract
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- Vibrio parahaemolyticus is a halophilic bacterium endemic to coastal areas, and its pathogenicity has caused widespread seafood poisoning. In our previous research, the protein expression of V. parahaemolyticus in Fe3+ medium was determined using isobaric tags for relative and absolute quantitation (iTRAQ). Here, nuclear magnetic resonance (NMR) was used to detect changes in the V. parahaemolyticus metabolome. NMR spectra were obtained using methanol-water extracts of intracellular metabolites from V. parahaemolyticus under various culture conditions, and 62 metabolites were identified, including serine, arginine, alanine, ornithine, tryptophan, glutamine, malate, NAD+, NADP+, oxypurinol, xanthosine, dCTP, uracil, thymine, hypoxanthine, and betaine. Among these, 21 metabolites were up-regulated after the stimulation of the cells by ferric iron, and 9 metabolites were down-regulated. These metabolites are involved in amino acid and protein synthesis, energy metabolism, DNA and RNA synthesis and osmolality. Based on these results, we conclude that Fe3+ influences the metabolite profiles of V. parahaemolyticus.
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Citations
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Frontiers in Microbiology.2018;[Epub] CrossRef
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- Spectral characterization of a pteridine derivative from cyanide-utilizing bacterium Bacillus subtilis - JN989651
- S. Durairaju Nisshanthini , Antony K. Teresa Infanta S. , Duraisamy Senthil Raja , Karuppannan Natarajan , M. Palaniswamy , Jayaraman Angayarkanni
- J. Microbiol. 2015;53(4):262-271. Published online March 4, 2015
- DOI: https://doi.org/10.1007/s12275-015-4138-0
- 309 View
- 0 Download
- 11 Crossref
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Abstract
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- Soil and water samples were collected from various regions of SIPCOT and nearby Vanappadi Lake, Ranipet, Tamilnadu, India. Based on their colony morphology and their stability during subculturing, 72 bacteria were isolated, of which 14 isolates were actinomycetes. Preliminary selection was carried out to exploit the ability of the microorganisms to utilize sodium cyanate as nitrogen source. Those organisms that were able to utilize cyanate were subjected to secondary screening viz., utilization of sodium cyanide as the nitrogen source. The oxygenolytic cleavage of cyanide is dependent on cyanide monooxygenase which obligately requires pterin cofactor for its activity. Based on this, the organisms capable of utilizing sodium cyanide were tested for the presence of pterin. Thin layer chromatography (TLC) of the cell extracts using n-butanol: 5 N glacial acetic acid (4:1) revealed that 10 out of 12 organisms that were able to utilize cyanide had the pterin-related blue fluorescent compound in the cell extract. The cell extracts of these 10 organisms were subjected to high performance thin layer chromatography (HPTLC) for further confirmation using a pterin standard. Based on the incubation period, cell biomass yield, peak height and area, strain VPW3 was selected and was identified as Bacillus subtilis. The Rf value of the cell extract was 0.73 which was consistent with the 0.74 Rf value of the pterin standard when scanned at 254 nm. The compound was extracted and purified by preparative High Performance Liquid Chromatography (HPLC). Characterization of the compound was performed by ultraviolet spectrum, fluorescence spectrum, Electrospray Ionization-Mass Spectrometry (ESI-MS), and Nuclear Magnetic Resonance spectroscopy (NMR). The compound is proposed to be 6-propionyl pterin (2-amino-6- propionyl-3H-pteridin-4-one).
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- Characterization of Exopolysaccharide (EPS) Produced by Weissella hellenica SKkimchi3 Isolated from Kimchi
- Min Ju Kim , Ha Na Seo , Tae Sik Hwang , Sung Hun Lee , Doo Hyun Park
- J. Microbiol. 2008;46(5):535-541. Published online October 31, 2008
- DOI: https://doi.org/10.1007/s12275-008-0134-y
- 204 View
- 0 Download
- 45 Crossref
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Abstract
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- Weissella hellenica SKkimchi3 produces the higher exopolysaccharide (EPS) on sucrose than lactose, glucose, and fructose at pH 5 and 20°C. Sucrose was exclusively used to cultivate SKkimchi3 in all experiments base on the EPS production tests. The molecular mass of EPS, as determined by gel permeation chromatography, was 203,000. 1H and 13C NMR analysis indicated that the identity of EPS may be a glucan. When EPS, starch, and cellulose was treated with α-amylase, glucoamylase, glucosidase, and cellulase, glucose was produced from starch and cellulose but was not produced from EPS. Based on HPLC analysis, elemental analysis, 1H and 13C NMR analysis, and enzymatic hydrolysis tests, EPS was estimated to be a glucan. EPS suspension was not precipitated even by centrifugation at 10,000×g for 60 min, and EPS made the fermented milk and bacterial culture viscous.
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- Characterization of C-P Lyase gene cluster by in vivo ³¹P-NMR spectroscopy
- Lee, Ki Sung , Kwak, In Young
- J. Microbiol. 1995;33(4):328-333.
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- ³¹P-NMR experiment was performed to detect phophonates (Pn) utilization and degradation in the several different C-P lyase mutants of E. coli and in E. aerogenes and the recombinants. The relative peak intensity (RPI) for the standard samples of 0.5 mM methylphosphonate (MPn) and 1.0 mM aminoethylphosphonate in glucose-MOPS medium showed 0.5 : 1.0 ratio. In the case of BW14329 (ΔphnC-P, ΔphoA), RPI did not change significantly after 24 hrs culturing, which means it nearly could not utilize Pn. In vivo ³¹P-NMR spectrum of E. aerogens (BWKL 16627) during 3 hrs starvation showed two intense peaks at 0-2 ppm and at near-10 ppm which indicate intracellular orthophosphate (Pi) and pyrophosphate (PPi), respectively. Both of them might be released by degradation of inorganic polyphosphate pool. When MPn is supplied to the medium as an unique P source, Pi content in the cell has the constant, but PPi seems to be slightly decreased. Recombinants (BWKL 16954) grew slower than E. aerogenes in the glucose-MOPS media with various P sources. In vivo ³¹P-NMR spectrum of recombinant did not show any intense signal in the cell. Surprisingly, under the cultivation adding with MPn, a few intense peaks in the region of Pi AND phospate monoester were detected.
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